Method for purifying antibodies

Patent metadata

Field	Value
Publication number	US-10906935-B2
Application number	US-201614995278-A
Country	US
Kind code	B2
Filing date	Jan 14, 2016
Priority date	Jul 25, 2014
Publication date	Feb 2, 2021
Grant date	Feb 2, 2021

Abstract

Disclosed here includes a method for purifying a biologic composition, comprising diafiltering the biologic composition into a composition comprising phosphate buffered saline (PBS) to obtain a purified composition. The method disclosed here can be particularly useful for removing one or more impurities from the biologic composition, such as bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane (Bis-tris).

First claim

What is claimed is:

1. A method for purifying a biologic composition, wherein the biologic composition comprises at least one impurity, wherein the impurity is bis ( 2 -hydroxyethyl)amino- tris(hydroxymethyl)methane (Bis-tris), wherein the method comprises diafiltering the biologic composition with phosphate buffered saline (PBS) to obtain a purified composition, wherein the biologic composition comprises a ch14.18 monoclonal antibody.

2. The method of claim 1 , wherein the ch14.18 monoclonal antibody is an isolated protein.

3. The method of claim 1 , wherein the concentration of the Bis-tris in the biologic composition is 10 to 50 mM Bis-tris at a pH of 6.3 to 6.7.

4. The method of claim 1 , wherein the diafiltering removes at least 50% of the Bis-tris from the biologic composition.

5. The method of claim 1 , wherein the diafiltering removes at least 70% of the Bis-tris from the biologic composition.

6. The method of claim 1 , wherein the concentration of the PBS is 10 to 50 mM Sodium Phosphate and 100 to 200 mM NaCl.

7. The method of claim 1 , wherein the ch14.18 antibody is concentrated to a concentration of at least 2.0 to 5.0 AU before being diafiltered into the composition comprising PBS.

8. The method of claim 1 , wherein the ch14.18 antibody is concentrated to a concentration of at least 4.0 to 6.0 AU before being diafiltered into the composition comprising PBS.

9. The method of claim 1 , further comprising isolating and purifying the ch14.18 antibody using at least one chromatography column.

10. The method of claim 9 , comprising isolating and purifying the ch14.18 antibody using at least one affinity chromatography column, at least one cation exchange chromatography column, and/or at least one anion exchange chromatography column.

11. The method of claim 10 , wherein the anion exchange chromatography column is a multimodal medium column.

12. The method of claim 11 , wherein the ch14.18 antibody is eluted from the multimodal medium column using a composition comprising Bis-tris.

13. The method of claim 1 , wherein at least three volume units of the biologic composition is diafiltered into one volume unit of the composition comprising PBS.

14. The method of claim 1 , wherein at least five volume units of the biologic composition is diafiltered into one volume unit of the composition comprising PBS.

15. The method of claim 1 , wherein the purified composition is further diafiltered into a composition comprising histidine.

16. A method for purifying a monoclonal antibody, comprising:
(a) passing a first composition comprising the monoclonal antibody through an affinity chromatography column to obtain a second composition comprising the monoclonal antibody;
(b) lowering the pH of the second composition to obtain a third composition;
(c) washing the third composition with a solvent-detergent to obtain a fourth composition;
(d) washing the fourth composition to remove the solvent-detergent to obtain a fifth composition;
(e) passing the fifth composition through a cation exchange chromatography column to obtain a sixth composition comprising the monoclonal antibody;
(f) subjecting the sixth composition to nano-filtration to obtain a seventh composition;
(g) passing the seventh composition through an anion exchange chromatography column to obtain an eighth composition comprising the monoclonal antibody and Bis-tris; and
(h) diafiltering the eighth composition into a composition comprising PBS to obtain a ninth composition comprising the monoclonal antibody but substantially free from Bis-tris.

17. A method for removing bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane (bis-tris) from a biologic composition, comprising diafiltering the biologic composition with phosphate buffered saline (PBS) to remove at least 50% of the bis-tris from the biologic composition, wherein the biologic composition comprises a ch14.18 monoclonal antibody.

Assignees

• United Therapeutics Corp
• Us Health

Inventors

• Meh David
• Atolagbe Timothy
• Farquharson G Mark
• Shaban Samir
• Koleck Mary
• Mitra George

Classifications

• C07K16/3084

  against tumour-associated gangliosides · CPC title

• C07K16/18

  against material from animals or humans · CPC title

• C07K1/36 Primary

  by a combination of two or more processes of different types · CPC title

• C07K1/34

  by filtration, ultrafiltration or reverse osmosis · CPC title

• B01D61/16

  Feed pretreatment · CPC title